Recombinant collagen production optimization in Escherichia coli
Thesis (M. Eng.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 2005.
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| Format: | Thesis |
| Language: | eng |
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Massachusetts Institute of Technology
2006
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| Online Access: | http://hdl.handle.net/1721.1/30974 |
| _version_ | 1811068938241441792 |
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| author | Whittemore, Brett A |
| author2 | Jean-François P. Hamel. |
| author_facet | Jean-François P. Hamel. Whittemore, Brett A |
| author_sort | Whittemore, Brett A |
| collection | MIT |
| description | Thesis (M. Eng.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 2005. |
| first_indexed | 2024-09-23T08:03:10Z |
| format | Thesis |
| id | mit-1721.1/30974 |
| institution | Massachusetts Institute of Technology |
| language | eng |
| last_indexed | 2024-09-23T08:03:10Z |
| publishDate | 2006 |
| publisher | Massachusetts Institute of Technology |
| record_format | dspace |
| spelling | mit-1721.1/309742022-01-13T07:54:29Z Recombinant collagen production optimization in Escherichia coli Whittemore, Brett A Jean-François P. Hamel. Massachusetts Institute of Technology. Dept. of Electrical Engineering and Computer Science. Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science Electrical Engineering and Computer Science. Thesis (M. Eng.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 2005. This electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections. Includes bibliographical references (p. 151-158). An Escherichia coli-based collagen-production process was used to investigate several process optimization objectives for use at the industrial scale. The effect of cooling on fermentation growth kinetics was studied, with preliminary results indicating that cooling an early to mid-batch phase fermentation (OD₆₀₀ = 2-4) to 10⁰C for up to 40 hours was not detrimental to the recovery of fermentation growth or the culture's ability to reach high cell density (OD₆₀₀> 50). A cost-effective assay of collagen-like polymer was examined under high-cell density conditions (as opposed to previously studied low density conditions) and an experimental design for tailoring the assay to high cell density fermentation samples is presented. In addition, a dual-plasmid strain of E. coli was designed for use in a novel process for the mass production of collagen-like polymers: one plasmid contains a thermally inducible recombinant collagen gene (CLP3.1-his), and the other contains an arabinose-inducible lytic gene (bacteriophage T4 t-holin) along with the basally expressed T7 lysozyme gene from the pLysS plasmid. A methodology for the optimization of the sequential induction of CLP3.1-his followed by induction of the t-holin is presented; a review of the literature suggests that decreased growth rate is detrimental to lytic efficiency (both the time required for lysis and the degree of lysis). The resulting process will enable lysis to take place in the bioreactor, thus avoiding the extra time and monetary cost of a separate cell homogenization step for E. coli disruption and endogenous protein release.. by Brett A. Whittemore. M.Eng. 2006-02-02T16:39:19Z 2006-02-02T16:39:19Z 2005 2005 Thesis http://hdl.handle.net/1721.1/30974 62560043 eng M.I.T. theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. See provided URL for inquiries about permission. http://dspace.mit.edu/handle/1721.1/7582 158 p. 9563685 bytes 9566105 bytes application/pdf application/pdf application/pdf Massachusetts Institute of Technology |
| spellingShingle | Electrical Engineering and Computer Science. Whittemore, Brett A Recombinant collagen production optimization in Escherichia coli |
| title | Recombinant collagen production optimization in Escherichia coli |
| title_full | Recombinant collagen production optimization in Escherichia coli |
| title_fullStr | Recombinant collagen production optimization in Escherichia coli |
| title_full_unstemmed | Recombinant collagen production optimization in Escherichia coli |
| title_short | Recombinant collagen production optimization in Escherichia coli |
| title_sort | recombinant collagen production optimization in escherichia coli |
| topic | Electrical Engineering and Computer Science. |
| url | http://hdl.handle.net/1721.1/30974 |
| work_keys_str_mv | AT whittemorebretta recombinantcollagenproductionoptimizationinescherichiacoli |